Open Access Highly Accessed Research article

Evolution of anterior Hox regulatory elements among chordates

Alfonso Natale1, Carrie Sims2, Maria L Chiusano3, Alessandro Amoroso1, Enrico D'Aniello16, Laura Fucci4, Robb Krumlauf25, Margherita Branno1 and Annamaria Locascio1*

Author affiliations

1 Laboratory of Cellular and Developmental Biology, Stazione Zoologica Anton Dohrn, Villa Comunale, Naples, Italy

2 Stowers Institute for Medical Research, Kansas City, MO 64110, USA

3 Department of Soil, Plant, Environmental and Animal Production Sciences, University of Naples "Federico II", Via Università 100, Portici, Italy

4 Department of Structural and Functional Biology, University of Naples "Federico II", Via Cinthia, Naples, Italy

5 Department of Anatomy and Cell Biology, Kansas University Medical School, Kansas City, KS 66160, USA

6 Molecular Cardiovascular Biology, MLC 7020 Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229, USA

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Citation and License

BMC Evolutionary Biology 2011, 11:330  doi:10.1186/1471-2148-11-330

Published: 15 November 2011



The Hox family of transcription factors has a fundamental role in segmentation pathways and axial patterning of embryonic development and their clustered organization is linked with the regulatory mechanisms governing their coordinated expression along embryonic axes. Among chordates, of particular interest are the Hox paralogous genes in groups 1-4 since their expression is coupled to the control of regional identity in the anterior nervous system, where the highest structural diversity is observed.


To investigate the degree of conservation in cis-regulatory components that form the basis of Hox expression in the anterior nervous system, we have used assays for transcriptional activity in ascidians and vertebrates to compare and contrast regulatory potential. We identified four regulatory sequences located near the CiHox1, CiHox2 and CiHox4 genes of the ascidian Ciona intestinalis which direct neural specific domains of expression. Using functional assays in Ciona and vertebrate embryos in combination with sequence analyses of enhancer fragments located in similar positions adjacent to Hox paralogy group genes, we compared the activity of these four Ciona cis-elements with a series of neural specific enhancers from the amphioxus Hox1-3 genes and from mouse Hox paralogous groups 1-4.


This analysis revealed that Kreisler and Krox20 dependent enhancers critical in segmental regulation of the hindbrain appear to be specific for the vertebrate lineage. In contrast, neural enhancers that function as Hox response elements through the action of Hox/Pbx binding motifs have been conserved during chordate evolution. The functional assays reveal that these Hox response cis-elements are recognized by the regulatory components of different and extant species. Together, our results indicate that during chordate evolution, cis-elements dependent upon Hox/Pbx regulatory complexes, are responsible for key aspects of segmental Hox expression in neural tissue and appeared with urochordates after cephalochordate divergence.